MSCs and meshes for Hernia - qPCR data.txt

Gene expression of cells infiltrated in the surgical meshes at 1 week and 1 month after surgical mesh implantation was analyzed by quantitative Real Time PCR (qPCR). qPCR was performed with commercial gene expression assays (Life Technologies, Thermo Fisher Scientific Inc.), using TaqMan Fast Advance Master Mix (Life Technologies, Thermo Fisher Scientific Inc.), in accordance with the protocol provided by the manufacturer. The amplification of cDNAs was performed in a QuantStudio 3 System (Applied Biosystems, Thermo Fisher Scientific) and the qPCR products were quantified by fluorescent method using the 2^-ΔCt expression. Duplicates of all samples were analyzed separately and normalized using HRPT1 as a reference gene.本研究通过实时定量聚合酶链式反应（quantitative Real Time PCR，qPCR），对手术补片植入1周及1个月后浸润至外科植入补片内的细胞的基因表达水平展开分析。实验采用赛默飞世尔科技（Thermo Fisher Scientific Inc.）旗下生命科技（Life Technologies）的商用基因表达检测试剂盒，搭配TaqMan Fast Advance预混试剂，严格遵循制造商提供的实验流程完成操作。互补脱氧核糖核酸（complementary DNA，cDNA）的扩增反应在QuantStudio 3系统（应用生物系统公司，赛默飞世尔科技）中进行，并通过荧光法结合2^-ΔCt表达量公式对qPCR产物完成定量分析。所有样本均设置复孔独立检测，并以HRPT1作为内参基因进行归一化处理。